Cancer immunotherapy aims to use and manipulate the host’s immune system to fight against cancer. The objective of this strategy is to induce specific and persistent immune responses leading to tumor eradication. Heat shock proteins (HSP) purified from cancer tissues have been identified as unique mediators of specific anti-tumor immunity. In our laboratory, we have developed an original vaccine, termed CRCL (Chaperone-Rich Cell Lysate) that consists of multiple HSP complexes enriched from tumor lysates. CRCL immunization leads to an efficient protection against a wide variety of murine cancers by inducing a strong, long-lasting, and specific T and NK-cell dependent immune responses against the tumor from which it has been generated. Tumor-derived CRCL has been shown to be more efficient in triggering DC activation than individual purified HSP or tumor lysates. The immunostimulatory effects of CRCL arise from its superior ability to provide a wide variety of tumor antigens to the immune system and by providing potent adjuvant effects. However, CD4⁺CD25⁺ regulatory T lymphocytes (Treg) critically contribute to the mechanisms of cancer-induced suppression. Data from independent groups including ours suggests they may also restrain the function of antigen presenting cells. The current study was designed to elucidate the molecular signaling events triggered by the tumor-derived CRCL vaccine in antigen presenting cells and evaluate whether CRCL may overcome the inhibitory effects of Treg modulation of DC and macrophage activation. Our results indicate CRCL activates DC and macrophages by inducing proinflammatory cytokine chemokine secretion. CRCL induces iNOS expression and NO production in macrophages. CRCL activation of DC and macrophages results in transcription factor NF-κB activation in vitro and in vivo, and this includes the activation of additional signaling molecules upstream of NF-κB. Following CRCL treatment the phenotypic maturation of DC, the production of DC and macrophage pro-inflammatory cytokines, and the activation of the transcription factor NF-κB are not affected by Treg. Additionally, CRCL induced activation of DC is not diminished by the immunosuppressive cytokine TGF-β 1. Our results indicate tumor-derived CRCL-treated DC and macrophages are refractory to Treg inhibition. These results are important for advancing CRCL-based vaccines in Phase I clinical trials.

Cancer immunotherapy aims to use and manipulate the host’s immune system to fight against cancer. The objective of this strategy is to induce specific and persistent immune responses leading to tumor eradication. Heat shock proteins (HSP) purified from cancer tissues have been identified as unique mediators of specific anti-tumor immunity. In our laboratory, we have developed an original vaccine, termed CRCL (Chaperone-Rich Cell Lysate) that consists of multiple HSP complexes enriched from tumor lysates. CRCL immunization leads to an efficient protection against a wide variety of murine cancers by inducing a strong, long-lasting, and specific T and NK-cell dependent immune responses against the tumor from which it has been generated. Tumor-derived CRCL has been shown to be more efficient in triggering DC activation than individual purified HSP or tumor lysates. The immunostimulatory effects of CRCL arise from its superior ability to provide a wide variety of tumor antigens to the immune system and by providing potent adjuvant effects. However, CD4⁺CD25⁺ regulatory T lymphocytes (Treg) critically contribute to the mechanisms of cancer-induced suppression. Data from independent groups including ours suggests they may also restrain the function of antigen presenting cells. The current study was designed to elucidate the molecular signaling events triggered by the tumor-derived CRCL vaccine in antigen presenting cells and evaluate whether CRCL may overcome the inhibitory effects of Treg modulation of DC and macrophage activation. Our results indicate CRCL activates DC and macrophages by inducing proinflammatory cytokine chemokine secretion. CRCL induces iNOS expression and NO production in macrophages. CRCL activation of DC and macrophages results in transcription factor NF-κB activation in vitro and in vivo, and this includes the activation of additional signaling molecules upstream of NF-κB. Following CRCL treatment the phenotypic maturation of DC, the production of DC and macrophage pro-inflammatory cytokines, and the activation of the transcription factor NF-κB are not affected by Treg. Additionally, CRCL induced activation of DC is not diminished by the immunosuppressive cytokine TGF-β 1. Our results indicate tumor-derived CRCL-treated DC and macrophages are refractory to Treg inhibition. These results are important for advancing CRCL-based vaccines in Phase I clinical trials.

en_US

dc.type

text

en_US

dc.type

Electronic Dissertation

en_US

dc.subject

Chaperone-Rich Cell Lysate

en_US

dc.subject

Dendritic Cells

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dc.subject

Macrophages

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thesis.degree.name

Ph.D.

en_US

thesis.degree.level

doctoral

en_US

thesis.degree.discipline

Cancer Biology

en_US

thesis.degree.discipline

Graduate College

en_US

thesis.degree.grantor

University of Arizona

en_US

dc.contributor.advisor

Katsanis, Emmanuel

en_US

dc.contributor.chair

Katsanis, Emmanuel

en_US

dc.contributor.committeemember

Bowden, George T.

en_US

dc.contributor.committeemember

Lybarger, Lonnie

en_US

dc.contributor.committeemember

Heimark, Ron

en_US

dc.contributor.committeemember

Larmonier, Nicolas

en_US

dc.identifier.proquest

10228

en_US

dc.identifier.oclc

659750825

en_US

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